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Validation of reference genes from Eucalyptus spp. under different stress conditions

DOI: 10.1186/1756-0500-5-634

Keywords: Eucalyptus, Drought, Cold, Light, Reference genes

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Eucalyptus globulus Labill, Eucalyptus urograndis (hybrid from Eucalyptus urophylla S.T. Blake X Eucalyptus grandis Hill ex-Maiden) and E. uroglobulus (hybrid from E. urograndis X E. globulus) were subjected to different treatments, including water deficiency and stress recovery, low temperatures, presence or absence of light, and their respective controls. Except for treatment with light, which examined the seedling hypocotyl or apical portion of the stem, the expression analyses were conducted in the apical and basal parts of the stem. To select the best pair of genes, the bioinformatics tools GeNorm and NormFinder were compared. Comprehensive analyses that did not differentiate between species, treatments, or tissue types, showed that IDH (isocitrate dehydrogenase), SAND (SAND protein), ACT (actin), and A-Tub (α-tubulin) genes were the most stable. IDH was the most stable gene in all of the treatments.Comparing these results with those of other studies on eucalyptus, we concluded that five genes are stable in different species and experimental conditions: IDH, SAND, ACT, A-Tub, and UBQ (ubiquitin). It is usually recommended a minimum of two reference genes is expression analysis; therefore, we propose that IDH and two others genes among the five identified genes in this study should be used as reference genes for a wide range of conditions in eucalyptus.The genus Eucalyptus belongs to the Myrtaceae family, which consists of approximately 600 species and subspecies, exhibits physiological plasticity with worldwide dispersion and successfully grows in different regions. Eucalyptus is a major source of cellulose for manufacturing paper [1] and globally, Brazil ranks first in the production of short fibre cellulose [2]. Due to its economic importance, the genome sequencing of the species Eucalyptus grandis was recently completed [3].E. globulus and E. grandis are the two main species cultivated in the world [4]. Among the existing species, E. globulus is known for it


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